Plasma Alkaline Phosphatase Activity Research Articles

Assessing safety, efficacy and residue depletion in golden mahseer, Tor putitora (Hamilton, 1822): biochemical and physiological responses to graded concentrations of oxytetracycline dietary supplementation.

The safety and effectiveness of oxytetracycline can potentially manage bacterial infections in fish. This, in turn, might reduce the concerns related to its use in aquaculture and human consumption, such as toxicity, antimicrobial resistance, and other associated risks. The primary objective of this study was to assess how adding oxytetracycline dihydrate to the diet affects its effectiveness, safety, and the presence of residues in T. putitora. T. putitora fingerlings, subjected to experimental infection with Aeromonas hydrophila at a concentration of 108 CFU mL- 1, received an oral administration of oxytetracycline dihydrate. The oxytetracycline dihydrate was added to the feed (corresponding to 2% of the fish body weight) at concentrations of 44.1, 88.2, 132.3 and 176.4mg Kg- 1 fish body weight per day. This treatment was carried out for 10 consecutive days. The biochemical and physiological responses of T. putitora and efficacy of oxytetracycline dihydrate were determined through estimation of microbial load (CFU mL- 1), haematogram, serum biomarkers, behavioral characteristics, non-specific immunity and residue depletion. Experimentally infected fish showed disease progression and induced histopathological conditions with highest microbial load (CFU mL- 1) in the muscle of both control and treated fish. The fish haematogram showed increased leucocyte and haemoglobin content, influenced by dietary oxytetracycline dihydrate. The fish demonstrated adaptive physiological response to oxytetracycline dihydrate at 44.1 to 88.2mg and resulted in increased albumin and globulin content. The serum-enzyme assay showed significant increase in aspartate aminotransferase (AST), alanine aminotransferase (ALT) and plasma alkaline phosphatase (ALP) activities in the test fish (< 0.05). Oxytetracycline dihydrate at 88.2 to 132.3mg Kg- 1 fish body weight per day recorded higher feed intake (75%), significant survivability (66-68%) and histopathological recovery. The suppressed immune response was manifested with decreased respiratory burst and lysozyme activity. The palatability, treatment of bacterial infection, histopathological changes and survivability by fingerlings of golden mahseer determined the safety and optimized the therapeutic potential of the oxytetracycline dihydrate at 88.2mg Kg- 1 fish body weight per day for 10 days to contain the infection by A. hydrophila. A withdrawal period of 8-d was recommended as oxytetracycline dihydrate concentration depleted below the legal maximum residue limit (MRL 2.0 mg g- 1) in the edible muscle of the golden mahseer reared at an average water temperature of 20°C. This is considered safe for human consumption.

Effects of Dietary Phytol Supplementation on Growth Performance, Immunological Parameters, Intestinal Bacteria, and Prevention of Oxidative Stress Following Transportation of Nile Tilapia, Oreochromis niloticus

Nile tilapia, Oreochromis niloticus, (2.00 ± 0.02 g) were reared in 16 70-L tanks (40 individual/tank) and fed diets (approx. 345 g/kg protein, approx. 87 g/kg crude fat) containing 0 (CTL), 75 (PH-75), 150 (PH-150), and 300 (PH-300) mg/kg phytol (n = 4). After 60 days of feeding (4% daily), growth performance, humoral immune parameters, and gut bacteria were analyzed. Also, hepatic antioxidant parameters were determined before and after the fish were transported in plastic bags for 6 hr. The results showed that PH-75 exhibited the highest final weight (P&lt;0.001), weight gain (P&lt;0.001), feed intake (P&lt;0.001), feed efficiency (P=0.015), plasma lysozyme activity (P=0.004), and intestinal Lactobacillus sp. population (P=0.017), among the treatments. The highest plasma alternative complement activity (P=0.006) and the lowest intestinal total viable bacteria (P=0.027) were observed in PH-75 and PH-150. The highest plasma alkaline phosphatase activities were observed in PH-75 and PH-300 (P=0.014). The highest blood leukocyte (P=0.008), monocyte (P=0.010), and eosinophil (P&lt;0.001) were observed in PH-300, while the highest blood neutrophil was observed in all phytol treatments (P&lt;0.001). The highest hepatic lipid peroxidation was observed in PH-300, whereas PH-75 and PH-150 showed the lowest values (P&lt;0.001). The highest hepatic reduced glutathione was observed in PH-75, also PH-150 exhibited significant elevation in this parameter, compared to CTL (P&lt;0.001). Transportation led to significant elevations in the hepatic antioxidant enzymes’ activities in CTL, PH-75, and PH-150; the highest activities were related to PH-75 and PH-150 treatments, which had also the highest post-transportation survivals (P&lt;0.001). In conclusion, phytol is a suitable feed supplement for Nile tilapia, improving growth performance and welfare, particularly at 75 mg/kg.

Influence of 19-Nortestosterone on the Content of Steroid Hormones, Hematological Indicators and Individual Links of Metabolism in Tissues of Carp (Cyprinus carpio)

The influence of different concentrations of the synthetic steroid 19-nortestosterone on the body mass and indexes of internal organs, hematological parameters, testosterone, progesterone and cortisol content, certain links of metabolic processes and enzymatic activity in the tissues of carp (&lt;i&gt;Cyprinus carpio&lt;/i&gt; L.) was studied. Under the short-term impact of 50 &amp;mu;g/dm&lt;sup&gt;3&lt;/sup&gt;, 19-nortestosterone increased the total protein and albumins content, creatine phosphokinase and aspartate aminotransferase activity, decreased the inorganic phosphorus and iron content, alkaline phosphatase and alanine aminotransferase activity in the blood plasma and did not affect the respiratory rate, external body features, the mass of hepatopancreas and spleen and their indexes, hematological indicators, the steroid hormones concentration in blood plasma and hepatopancreas, content of glucose, triglycerides, creatinine, calcium, inorganic phosphorus in blood plasma. Under 200 &amp;mu;g/dm&lt;sup&gt;3&lt;/sup&gt;, the respiratory rate and ESR increased, as well as the cortisol and testosterone content in blood plasma and hepatopancreas, whereas progesterone in blood plasma decreased and in hepatopancreas increased. High concentration of 19-nortestosterone caused elevated protein and albumin content, creatine phosphokinase and aspartate aminotransferase in the blood plasma activity, whereas glucose, inorganic phosphorus, iron, alkaline phosphatase activity was lower. The 19-nortestosterone did not affect the mass and index of hepatopancreas and spleen, hemoglobin concentration, the erythrocytes, leukocytes and lymphocytes number and ratio of eosinophils, neutrophils, lymphocytes and monocytes, as well as triglycerides, creatinine, calcium content, lactate dehydrogenase and alanine aminotransferase activity in blood plasma. The conclusion was made about the carp's ability to adapt to low concentrations of the synthetic steroid 19-nortestosterone under its short-term impact by modification of the certain links of metabolism and enzymatic activity. High concentration of 19-nortestosterone significantly affected the hormonal status and metabolism of carp.

Effects of dietary Radix bupleuri root extract on the growth, muscle composition, histology, immune responses and microcystin-LR stress resistance of juvenile red claw crayfish (Cherax quadricarinatus)

The current study was conducted to determine the effects of dietary Radix bupleuri root extract (RBE) on the growth performance, muscle composition, immune responses, and microcystin-LR (MC-LR) stress resistance of juvenile red claw crayfish (Cherax quadricarinatus). Crayfish (0.28 ± 0.02 g) were fed experimental diets containing graded RBE levels as 0 (Diet 1), 0.2 (Diet 2), 0.4 (Diet 3), 0.6 (Diet 4), 0.8 (Diet 5) and 1.6 (Diet 6) g/kg RBE for 8 weeks. The results showed that specific growth rate (SGR) and weight gain rate (WGR) were significantly increased in the Diet 4 and Diet 5 groups. The content of muscle crude protein was significantly increased in the Diet 4 and Diet 5 groups, and the crude lipid content was significantly decreased in the Diet 5 group. Dietary RBE did not alter the structure of the hepatopancreas. Total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) activities in hepatopancreas and plasma were significantly increased in Diet 4 and Diet 5 groups compared to the control group, while the malondialdehyde (MDA) content in hepatopancreas and plasma was significantly decreased in the Diet 5 group. Plasma alkaline phosphatase (AKP) activity was significantly higher in the Diet 3, Diet 4, Diet 5 and Diet 6 groups. Hepatopancreas and plasma PO activities were significantly increased in Diet 4 and Diet 5 groups. The mRNA levels of extracellular copper/zinc superoxide dismutase (ecCu, Zn-SOD), hemocyanin, C-type lysozyme (C-LZM), GPx, and selenium-dependent glutathione peroxidase (Se-GPx) were significantly up-regulated in the hepatopancreas of crayfish fed Diet 5 than those of the control group. After MC-LR challenge, the survival rates of crayfish in the Diet 4 and Diet 5 groups were significantly higher than the control group. The findings suggest that 0.6–0.8 g/kg of RBE can improve growth and muscle composition and enhance immunity and microcystin-LR resistance.

313 Effect of Dietary Trace Mineral Source on Performance, Bones Mineralization and Mineral Status in Fattening Pigs

Abstract Several papers report greater bioavailability of dietary trace mineral fed as organic in pigs compared with inorganic sources. However, only a few studies have tried to assess the variability of the trace mineral bioavailability among different organic sources. Growing barrows (n = 54; initial BW = 28.9 ± 1.3 kg) were used to determine the effects of trace mineral source on growth performance, bones mineralization and mineral status over the entire fattening period (d0-d73). Pigs were blocked by BW and randomly assigned to supplementation of trace minerals as inorganic (ITM) or one of 2 organic sources (OTM): glycine complex (GLY) or amino acids complex (AA). During the two feeding phases (growing: d0-d35; finishing: d35-d73), the experimental diets were supplemented with 5, 35 and 5 mg/kg of Cu, Zn and Mn, respectively. Pigs were fed restricted all through the experimental period. Feed intake was therefore comparable all through the experimental period for all pigs. Growth performance did not differ among the three experimental groups during the growing phase. However, Feed:Gain ratio measured during the finishing period tended to be affected by the dietary treatments (P=0.10). When grouping the GLY and AA pigs, the Feed:Gain ratio measured from d35 to d73 for the OTM pigs was significantly lower compared with ITM pigs (-0.07, P = 0.03). There was no significant effect of the dietary treatment on plasma trace mineral concentrations and alkaline phosphatase activity measured at day 34. At the end of the growing phase, plasma ceruloplasmin activity tended to be greater for the GLY and AA groups in comparison with the ITM pigs (+6.9%, P = 0.06). The zinc deposition in the proximal phalanx was significantly increased when the trace minerals were offered as organic compared with the group ITM (+10.9%, P = 0.06). Zinc content in the liver was greater for the pigs fed the OTM sources compared with the others (+6.0%, P = 0.10). The quantity of copper deposited in the liver did not differ significantly among the 3 trace mineral sources. This study indicates that the total replacement of inorganic Zn, Cu and Mn by organic sources brings benefits in terms of feed efficiency in fattening pigs. A better bioavailability as suggested by greater deposition levels in bone and liver as well as the increased circulating ceruloplasmin activity could explain the improvement of performance during the finishing period. Regardless of the variable considered, no significant differences were noticed between the two organic trace mineral sources.

Effects of two phosphorous sources in the diet on the growth performance, digestibility, and plasma physiological parameters of Pelodiscus sinensis juveniles

Phosphorus is an essential mineral for aquatic animals to maintain the health of the skeletal system and many physiological functions. This study assessed the effects of two inorganic phosphorus sources on growth performance, apparent phosphorus digestibility, whole-body proximate composition, and physiological status in juvenile Pelodiscus sinensis. Two experimental diets were supplemented with 4% calcium phosphate monobasic (MCP) and 5.47% calcium phosphate dibasic (DCP), respectively, to obtain equal total dietary phosphorus (2.20%). 96 turtles (initial body weight: 5.40±0.03g) were randomly distributed into 12 tanks and fed the corresponding diets for 60d. Results showed that phosphorus sources have not significantly influenced the growth parameters, including the specific growth rate, feeding rate, and feed conversion ratio (P&gt;0.05). No significant differences were observed in the hepatosomatic index and whole-body proximate compositions between MCP and DCP groups (P&gt;0.05). The apparent digestibility coefficients of dry matter and phosphorus in MCP group (53.22%) are slightly higher than that in DCP group (48.98%) but did not reach the statistically significant level (P &gt; 0.05). Turtles in MCP and DCP groups are the same in plasma physiological parameters and have equal alkaline phosphatase activities in plasma and liver (P&gt;0.05). In conclusion, calcium phosphate monobasic and calcium phosphate dibasic had the same biological phosphorus availability in diet for juvenile Pelodiscus sinensis.

Impact of zinc, copper, manganese and chromium supplementation on growth performance and blood metabolic profile of Sahiwal (Bos indicus) male calves.

This study aimed to see the effect of oral supplementation of specific trace minerals mixture on the growth, immunity, and reproductive development of indigenous growing bull calves. Eighteen Sahiwal bull calves, with an average age of 6 months were chosen and divided into three groups. Group 1 was fed with a basal diet, Group 2 was provided with an additional specific trace mineral supplement to achieve a diet containing 70 ppm of Zn, 17.50 ppm of Cu, 65 ppm of Mn, and 1.75 ppm of Cr. Group 3 received a 25% extra supplement to achieve a diet containing 87.50 ppm of Zn, 21.87 ppm of Cu, 81.25 ppm of Mn, and 2.18 ppm of Cr. The experiment was carried out for a total of 180 days. According to the findings, there was no significant impact of specific trace minerals supplementation on the animals' body weight, morphometric parameters, dry matter intake, average daily gain, nutritional value, digestibility and nitrogen retention. However, higher levels of Zn, Cu, and Mn led to increased (p < 0.05) total retention, while Cr retention remained the same. Serum mineral concentrations of Zn, Cu, and Mn increased significantly (p < 0.05) in G2 and G3 compared to the G1 group while Ca, P, and Cr had no significant change. Blood plasma glucose, albumin, globulin, and total protein showed no significant differences. Plasma alkaline phosphatase activity improved significantly (p < 0.05) in G2 and G3 but alanine transaminase, aspartate aminotransferase, hemoglobin, hematocrit, and IGF-1 remained unchanged. Superoxide dismutase activity, ferric-reducing antioxidant power, and total immunoglobulin concentration increased significantly (p < 0.05) in G2 and G3 groups, however, catalase activity and IgG count did not change among the groups. Mineral-supplemented groups (G2 and G3) showed a significant change (p < 0.05) in testosterone production during the 120th and the 180th day of the trial. Scrotal circumference and temperature gradient of the scrotal surface did not show any significant change. Supplementing growing bull calves with specific trace minerals above the basal level (70, 17.50, 65 and 1.75 ppm of Zn, Cu, Mn and Cr) has no direct beneficial effect on the growth parameters but can have positive effects on their antioxidant status, immunity and reproductive development as the related blood parameters were positively affected.

Prebiotics Together with Raspberry Polyphenolic Extract Mitigate the Development of Nonalcoholic Fatty Liver Diseases in Zucker Rats.

Previous studies suggested that dietary supplementation with prebiotic fructooligosaccharides (FOSs) and polyphenols could mitigate disorders related to the first stage of nonalcoholic fatty liver disease (NAFLD) induced by an obesogenic diet. Therefore, this experiment aimed to address whether the health-promoting potential of raspberry polyphenols together with FOSs can regulate advanced-stage NAFLD in Zucker rats genetically predisposed to develop obesity. The addition of FOSs and raspberry polyphenolic extract to the diet reduced liver fat accumulation and triglyceride, free fatty acid, and total cholesterol levels in the liver. The elevated GSH/GSSG ratio and reduced malondialdehyde content indicated that the liver antioxidant potential was considerably increased. The treatment also lowered the plasma aminotransferase and alkaline phosphatase activities and collagen type IV levels. Insulin levels were decreased, but glucose levels remained constant, indicating greater insulin sensitivity. These changes may result from the upregulation of FXR and AHR receptors in the liver, which are responsible for regulating lipid metabolism and glucose and bile acid synthesis. The reduced bile acid levels in the cecal contents confirmed the activation of liver mechanisms. In conclusion, dietary enrichment with FOSs and raspberry polyphenolic extract has sufficient health-promoting potential to regulate liver metabolism, oxidative stress, and inflammation related to NAFLD development in obese Zucker rats.

Biochemical Response and Histopathologic Alterations in Papyrocranus afer from Banegbe River, Delta State, Nigeria

Aim: This study investigated the effects of pollutants on the biochemical and histopathological components of Papyrocranus afer obtained from the Banegbe River at different locations (upstream, middle-stream, and downstream). The effect of the pollutant on the checked were, levels of alkaline phosphatase (ALP), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) in blood samples of the fishes and the histopathologic alterations of the gills and liver of the fishes. &#x0D; Methodology: The gills and liver tissue of the fish were collected and processed using routine histologic techniques for fixing and embedding in paraffin and staining sections with hem eosin. All the indices of the enzyme tests were estimated with the aid of Randox Diagnostic Test Kits following the manufacturer’s methods. &#x0D; Results: Evaluation of the plasma enzymes showed significant increases (p&lt;0.05) in plasma ALT, AST, and ALP activity in all the sampled points of Banegbe River when compared to the control 1 (Orogodo river) with middle-stream having significant (p&lt;0.05) higher concentrations when compared to down-stream and up-stream. The main alterations observed in the liver of the fish at the Banegbe River were Kupffer cell hyperplasia and focal necrosis of the liver. While the gills showed alteration of the structure of the epithelium, hyperplasia of the epithelium of the primary lamella, epithelial lifting, vacuolization, alteration of the structure, and occurrence of aneurysms in the secondary lamella, all these alterations were as a result of pollutant in the river. Control 1 showed normal parenchymal cells of the liver and normal gill structure. &#x0D; Conclusion: The increase in enzyme activities suggests leakage of these enzymes from the liver cytosol into the bloodstream as a result of liver damage by contaminants. From our results, it can be concluded that the Banegbe River is polluted as a result of effluents channeled in the river by industries within the environs.

Hepatic Health and Humoral Immunological Parameters of Common Carp (Cyprinus carpio) Fed Lactic Acid-supplemented Diets

Background: Organic acids and their salts are known as appropriate substitutes in feed for improving the health, growth, and performance of fish. Objectives: The present study was conducted to determine the effects of dietary lactic acid supplementation on immunological factors, hepatic enzyme activity, and plasma proteins in common carp, Cyprinus carpio. The fish were fed the diets mentioned above for 56 days, then their growth performance, humoral immunity, and plasma hepatic enzymes were assessed. Methods: A total of 180 fish (mean weight=25 g) were randomly distributed in twelve tanks (150 L water in each tank) as four treatments, fed diets containing 0, 2.5, 5, and 10 g/kg lactic acid (T0, T1, T2, and T3, respectively). Results: At the end of the feeding trial, T2 showed significantly higher growth performance than T0. Plasma total protein, albumin, and globulin levels of T1-T3 were significantly higher than that of T0. However, plasma protein levels decreased significantly by elevation in dietary lactic acid concentration (10 g/kg). No significant differences were observed in plasma aspartate aminotransferase (AST) and alkaline phosphatase (ALP) activity among the treatments, although alanine aminotransferase (ALT) activity decreased significantly in fish-fed dietary lactic acid supplementation (T1-T3) compared with the control group (T0). All humoral immunity parameters (lysozyme, complement, immunoglobulin, and bactericidal activity) increased significantly in T1-T3 treatments compared to the T0 group. Conclusion: Overall, dietary lactic acid supplementation improves growth performance, humoral immunological parameters, and hepatic health. According to the results, dietary lactic acid (2.5-5 g/kg) is recommended for preparing common carp feed.

Selenium-Enriched Cardamine violifolia Alleviates LPS-Induced Hepatic Damage and Inflammation by Suppressing TLR4/NODs-Necroptosis Signal Axes in Piglets.

Selenium-enriched Cardamine violifolia (SEC), a cruciferous plant, exerts excellent antioxidant and anti-inflammatory capacity, but its effect on hepatic function is unclear. This study investigated the effect and potential mechanism of SEC on hepatic injury induced by lipopolysaccharide (LPS). Twenty-four weaned piglets were randomly allotted to treatment with SEC (0.3 mg/kgSe) and/or LPS (100 μg/kg). After 28 days of the trial, pigs were injected with LPS to induce hepatic injury. These results indicated that SEC supplementation attenuated LPS-induced hepatic morphological injury and reduced aspartate aminotransferase (AST) and alkaline phosphatase (ALP) activities in plasma. SEC also inhibited the expression of pro-inflammatory cytokines such as interleukin 6 (IL-6) and tumor necrosis factor-alpha (TNF-α) after the LPS challenge. In addition, SEC improved hepatic antioxidant capacity via enhancing glutathione peroxidase (GSH-Px) activity and decreasing malondialdehyde (MDA) concentration. Moreover, SEC downregulated the mRNA expression of hepatic myeloid differentiation factor 88 (MyD88) and nucleotide-binding oligomerization domain proteins 1 (NOD1) and its adaptor molecule receptor interacting protein kinase 2 (RIPK2). SEC also alleviated LPS-induced hepatic necroptosis by inhibiting RIPK1, RIPK3, and mixed-lineage kinase domain-like (MLKL) expression. These data suggest that SEC potentially mitigates LPS-induced hepatic injury via inhibiting Toll-like receptor 4 (TLR4)/NOD2 and necroptosis signaling pathways in weaned piglets.

Effects of different dietary lipid sources (perilla, fish, and soybean oils) on growth, lipid metabolism, antioxidant, and immune status in Chinese giant salamander (Andrias davidianus)

IntroductionPerilla oil is a natural vegetable oil rich in alpha-linolenic acid extracted from Perilla seeds. Fish oil is a commonly used animal fat containing a large amount of n-3 polyunsaturated fatty acids. Soybean oil is a commonly used vegetable oil rich in linoleic acid. This study aimed to determine whether perilla, fish, or soybean oil can be used as a lipid source in the diet of the Chinese giant salamander (Andrias davidianus).MethodsThree isonitrogenous and isolipidic diets were created by adding 3.0% each of soybean oil (SO), fish oil (FO), and perilla seed oil (PSO). Eight adolescent giant salamanders were fed in rectangular tanks for 98 days with a total of 96 giant salamanders (mean body weight = 75.20g) and were fed an experimental diet until satiation.Results and DiscussionThe viscerosomatic index and crude lipid content of muscle were found to be considerably lower and the muscle crude protein was substantially higher in the PSO group than in the other groups (P&amp;lt;0.05). Linoleic acid levels were highest in SO, eicosapentaenoic acid and docosahexaenoic acid levels were highest in FO, and α-linolenic acid levels were highest in PSO in A. davidianus muscle tissue (P&amp;lt;0.05). The results of study indicate that the PSO diet can significantly increase the total antioxidative capabilities in the liver and intestine, by elevating the activities of total superoxide dismutase, carnitine transferase-1, and acetyl-CoA carboxylase in the liver. Meanwhile, the immunoglobulin M and high-density lipoprotein cholesterol levels were higher in the blood (P&amp;lt;0.05). Furthermore, it significantly reduced hepatic malondialdehyde, plasma endotoxin, D-lactic acid, and total cholesterol levels, and plasma alkaline phosphatase, diamine oxidase, aspartate transaminase, and alanine transaminase activities (P&amp;lt;0.05). Therefore, perilla, fish, and soybean oils can be used as single lipid sources for A. davidianus with respect to growth performance. On the other hand, perilla oil can enhance crude protein content of muscle, increase Immunoglobulin M(IgM) content and Alkaline phosphatase(AKP) enzyme activity, promote lipid metabolism, and maintain the health of the liver and intestine in A. davidianus.

Cadmium accelerates autophagy of osteocytes by inhibiting the PI3K/AKT/mTOR signaling pathway.

Cadmium (Cd) can damage bone cells and cause osteoporosis. Osteocytes are the most numerous bone cells and also important target cells for Cd-induced osteotoxic damage. Autophagy plays important role in the progression of osteoporosis. However, osteocyte autophagy in Cd-induced bone injury is not well characterized. Thus, we established a Cd-induced bone injury model in BALB/c mice and a cellular damage model in MLO-Y4 cells. Aqueous Cd exposure for 16 months showed an increase in plasma alkaline phosphatase (ALP) activity and increase in urine calcium (Ca) and phosphorus (P) concentrations in vivo. Moreover, expression level of autophagy-related microtubule-associated protein 1A/1B-light chain 3 II (LC3II) and autophagy-related 5 (ATG5) proteins were induced, and the expression of sequestosome-1 (p62) was reduced, along with Cd-induced trabecular bone damage. In addition, Cd inhibited the phosphorylation of mammalian target of rapamycin (mTOR), protein kinase B (AKT), and phosphatidylinositol 3-kinase (PI3K). In vitro, 80 μM Cd concentrations exposure upregulated LC3II protein expression, and downregulated of p62 protein expression. Similarly, we found that treatment with 80 μM Cd resulted in a reduction in the phosphorylation levels of mTOR, AKT, and PI3K. Further experiments revealed that addition of rapamycin, an autophagy inducer, enhanced autophagy and alleviated the Cd-induced damage to MLO-Y4 cells. The findings of our study reveal for the first time that Cd causes damage to both bone and osteocytes, as well as induces autophagy in osteocytes and inhibits PI3K/AKT/mTOR signaling, which could be a protective mechanism against Cd-induced bone injury.

Supplemental Microbial Phytase Improves Bioavailability of Dietary Zinc to Weanling Pigs

Two experiments were conducted to determine the effects of supplemental microbial phytase on utilization of dietary zinc by weanling pigs. Experiment 1 was a 2 x 3 factorial arrangement of treatments with 24 pigs for 4 wk. Two levels of phytase activity (0 and 1350 units/g) and three levels of zinc (0, 30 and 60 mg/kg as ZnSO4.7H2O) were added to a corn-soybean meal basal diet. Weekly measures included growth performance, plasma alkaline phosphatase activity and plasma mineral concentrations. In Experiment 2, mineral balances were determined in 12 pigs fed the basal diet or the diet with added zinc (30 mg/kg) or phytase (1350 units/g). The results indicated that either supplemental phytase or supplemental zinc increased plasma alkaline phosphatase activity and plasma zinc concentrations, but these increases were not additive. Supplemental phytase decreased plasma alkaline phosphatase activity in pigs supplemented with zinc. Supplemental phytase also significantly enhanced weight gain, feed intake, gain:feed ratio, plasma concentrations of inorganic phosphorus, and retention of phosphorus and calcium. Neither supplemental phytase nor zinc affected zinc retention. Supplementing corn-soybean meal diets with microbial phytase at 1350 units/g feed improves bioavailability of zinc as well as of phytate phosphorus to weanling pigs.

Dietary Fat Exacerbates Liver Disease in Bile Duct-Ligated Rats

Previous studies have suggested that the mortality of bile duct-ligated (BDL) rats is related to the amount of dietary fat consumed. We investigated the influence of dietary fat concentration on liver disease in BDL rats. Groups of rats were fed for 4 wk either a low fat diet (LF, 0.92 kJ/g; 3% of total energy from fat), a high fat diet (HF, 1.07 kJ/g; 30% fat), a high fat diet with energy density equivalent to that of the LF diet (HFIB, 0.92 kJ/g; 30% fat) or a diet based on the composition of commercial nonpurified diets (COMP, 0.90 kJ/g; 10% fat). Energy intake, body weight gain, plasma biochemical indices and hepatic histology were compared in BDL and sham-operated control rats. Bile duct-ligated animals consuming the LF diet showed a faster recovery of energy intake and greater body weight gain following surgery than did BDL animals fed the other three diets. Plasma alkaline phosphatase activity was significantly greater in BDL animals fed either of the high fat diets than in those fed the LF or COMP diet starting 1 and 2 wk, respectively, after surgery. Hepatic fibrosis and bile duct proliferation at d 28 post-ligation were greater in rats fed a 30% fat (HF) diet than in rats fed the 3% fat (LF) diet. These results suggest that dietary fat concentration can influence the severity of liver dysfunction in extrahepatic biliary obstruction.

Evaluation of Cottonseed Meal as an Alternative to Fish Meal in Diet for Juvenile Asian Red-Tailed Catfish Hemibagrus wyckioides.

A 10-week trial was performed to investigate the effects of replacing fishmeal with cottonseed meal (CSM) on the growth rate, protein metabolism, and antioxidant response of Asian red-tailed catfish Hemibagrus wyckioides. Five isonitrogenous and isocaloric diets (C0, C8.5, C17.2, C25.7, and C34.4) were prepared to contain 0%, 8.5%, 17.2%, 25.7%, and 34.4% CSM replacing fishmeal, respectively. The weight gain, daily growth coefficient, pepsin, and intestinal amylase activities initially increased and then decreased with the raising dietary CSM levels; the highest values were observed in the C17.2 group (P < 0.05). However, feed cost exhibited the opposite trend. With the increasing dietary CSM levels, the protein efficiency ratio and intestinal trypsin activity decreased but feed conversion rate increased gradually; while no differences were observed among the C0, C8.5, and C17.2 groups (P > 0.05). Dietary CSM inclusion regardless of levels increased the plasma growth hormone level as well as hepatic aspartate aminotransferase (AST) and γ-glutamyl transpeptidase activities but decreased the plasma glutamate dehydrogenase and AST activities (P < 0.05). With the increasing dietary CSM levels, the plasma alkaline phosphatase (AKP) and hepatic superoxide dismutase activities decreased but malondialdehyde content increased gradually, while no differences were observed among the C0, C8.5, and C17.2 groups (P > 0.05). The plasma immunoglobulin M content and hepatic glutathione reductase activity initially increased but then decreased with the raising dietary CSM levels; the highest values were found in the C17.2 group. These results indicated that dietary CSM inclusion level up to 17.2% improved the growth rate, feed cost, digestive enzyme activity, and protein metabolism without compromising antioxidant capacity of H. wyckioide, whereas these parameters were depressed by further inclusion of CSM. CSM is a potentially cost-effective alternative plant protein source in diet of H. wyckioide.

Effects of triterpenoids in fatty products on liver condition of laboratory animals with acute toxic hepatitis

One of the principles of prevention and non-medicamentous treatment of liver diseases, including hepatitis of different etiology, is the normalization of the diet through the consumption of food with physiologically active ingredients, in particular betulin, which helps to eliminate the causes of metabolic and oxidative disorders within liver cells. The aim of the research was to assess in vivo the influence of triterpene alcohol betulin extracted from Betula pendula Roth. birch bark in fat-containing products (for example mayonnaise) on the blood biochemical parameters and liver morphological structure of rats with initiated acute toxic hepatitis. Material and methods. Hepatoprotective and antioxidant activities of betulin as part of mayonnaise samples has been investigated in vivo on the model of toxic hepatitis initiated by carbon tetrachloride in male Wistar rats weighing 210-265 g. The animals were divided into 4 groups of 10 animals each: CG-1 - intact, CG-2 and MG - with carbon tetrachloride initiated toxic hepatitis. rats of the main groups were orally administered mayonnaise once a day at a dosage of 1 ml for 21 days after the formation of the model pathology: OG-1 with the added betulin (1 mg per 1 kg of body weight), OG-2 without betulin. Disorders of metabolic and oxidative processes in liver cells of animals were evaluated by biochemical indicators of blood plasma: the level of glucose, albumin, total cholesterol, triglycerides and urea and the activity of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and γ-glutamyltransferase. Oxidative stress in rats was estimated by the activity of catalase and superoxide dismutase in blood hemolysate (at a dilution of 1:200 and 1:10, respectively); the total prooxidant (in blood plasma) and total antioxidant (in blood hemolysate at a dilution of 1:10) activity were determined spectrophotometrically (colored complexes of TWIN-80 oxidation products with thiobarbituric acid). The morphological structure of rats' liver was estimated by microscopy of prepared cuts of hepatic tissue. Results. Based on biochemical parameters of rat blood plasma, it has been established that the administration of mayonnaise with betulin prevents the development of cytolic syndrome and suppresses the process of peroxidation by directly neutralizing free radicals. Aspartate aminotransferase and alkaline phosphatase activity in blood plasma of the experimental animals of the main group MG-1 reduced by 20.7 and 35.2% compared with indicators of the rats of the main group MG-2. Glucose concentration normalized to the level of the control group CG-1. The concentration of bilirubin and triglycerides decreased by 22.9 and by 48.1%, which indicates a significant reduction in the indicators of cholestatic syndrome in the group of animals OG-1 compared to OG-2. The total prooxidant activity and the concentration of thiobarbiturate-reactive products decreased compared to the CG-2 and MG-2 groups, which indicates the suppression of oxidative stress and, as a result, an improvement in liver conditions of animals with toxic hepatitis even when taking a fat-containing product. In liver histopeparates of animals receiving mayonnaise with betulin, necrobotic changes were less pronounced in comparison with the group MG-2. They were estimated at 1 point: small-drip dystrophy spots were found, haemorrhages in the interregional septum with inflammatory infiltration in the course of hemorrhages against the presence of necrosis hepatocytes with pronounced adipose dystrophy in the centres of the lobules, step necrosis with signs of replacing the damaged hepatocytes of the connective tissue, accompanied by centrolobular hemorrhages in MG-2 rats. Conclusion. Introduced into the composition of mayonnaise betulin, reduces the development of cytolic syndrome in toxic hepatitis and suppresses the process of peroxidation, on the basis of which fat-containing foods with betulin can be recommended for clinical examination as specialized products in acute and chronic liver diseases, including complicated cholestasis.

ENZYME ACTIVITY OF CARBOHYDRATE AND AMINOACID METABOLISM, LIPID PEROXIDATION IN CARP TISSUES UNDER THE EFFECT OF 19-NORTESTOSTERONE

The aim of the study was to investigate the activity of a number of enzymes of carbohydrate metabolism, individual amino acids, creatine phosphate, inorganic phosphorus and lipid peroxidation (LPO) in the hepatopancreas and blood plasma of carp (Cyprinus carpio L.) under the influence of the synthetic steroid 19-norethisterone. The experiments were conducted on two-year-old carps, which were kept for 24 hours in aquariums with different concentrations of 19-nortestosterone in water at optimal temperature, oxygen content and pH. The activity of lactate dehydrogenase (LDH), alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatine phosphokinase (CPK) in the supernatant fraction of hepatopancreatic homogenate was monitored by UV-kinetic and colourimetric methods. The activity of isocitrate dehydrogenase (ICDH) was determined spectrophotometrically by the accumulation of NADPH, and lactate dehydrogenase (LDH), glucose-6-phosphate dehydrogenase (Gl-6-PDH) and glucose-6-phosphatase (Gl-6-Phase) - by the accumulation of NADH in the reactions of conversion of the corresponding substrates. Conventional biochemical methods were used to determine the concentration of diene conjugates, malondialdehyde (MDA), lipid hydroperoxides, catalase (CAT) and superoxide dismutase (SOD) activity in blood plasma and supernatant fraction of hepatopancreatic homogenate. Conventional biochemical methods were used to determine the concentration of diene conjugates, malondialdehyde (MDA), lipid hydroperoxides, catalase (CAT) and superoxide dismutase (SOD) activity in blood plasma and the supernatant fraction of hepatopancreatic homogenate. It was found that fish exposed to water with a concentration of synthetic steroid 19-nortestosterone of 50 µg/dm3 for 24 hours had higher levels of MDA and diene conjugates, increased activity of CAT and SOD in blood plasma, increased activity of ALP, ALT, AST, CK, CAT and SOD in the hepatopancreatic supernatant fraction, and LDH remained unchanged, as well as the level of lipid hydroperoxides in blood plasma. Under the influence of 19-nortestosterone, the activity of Gl-6-FDG and IDDG in the mitochondrial fraction of carp hepatopancreas increases, and that of Gl-6-phase in the supernatant. At a concentration of 19-nortestosterone in water of 200 μg/dm3, the content of lipid hydroperoxides in the blood plasma of carp increases, and in the hepatopancreatic supernatant fraction - diene conjugates and SOD activity, while ALP and CPK decrease relative to their values in fish with a low level of xenobiotic in water. The content of MDA, diene conjugates, as well as the activity of CAT, SOD, ALP, CPK and LDH in blood plasma, IDH and Gl-6-PDH in the mitochondrial fraction and Gl-6-Phase and LDH in the hepatopancreatic supernatant did not change, but remained higher than in the control. It was concluded that 19-nortestosterone stimulates carbohydrate metabolism, amino acids alanine and aspartate transamination, creatine phosphate conversion, antioxidant enzyme activity and lipid peroxidation in the hepatopancreas of two-year-old carp. The obtained results deepen the understanding of the mechanism of action of synthetic steroids in bony fish, and the differences in enzyme activity and lipid peroxidation can be important criteria for ecotoxicological assessment of water bodies contaminated with hormonal compounds.

Fluorescence Turn-on Detection of Alkaline Phosphatase Activity and Al3+ Using Vitamin B6 Cofactor Conjugated GSH Capped Mn-doped ZnS Quantum Dots.

The glutathione (GSH) functionalized Mn-doped ZnS quantum dots (GSH_Mn_ZnS QDs) was conjugated with pyridoxal 5'-phosphate (PLP). The -CHO group of vitamin B6 cofactor PLP interacted with the -NH2 group of GSH functionalized Mn_ZnS QDs. The conjugation of PLP quenched the fluorescence emission of GSH_Mn_ZnS QDs at 601nm. Addition of alkaline phosphatase (ALP) catalytically dephosphorylated the PLP into pyridoxal that restored the fluorescence emission of GSH_Mn_ZnS QDs. With a sensitivity of 0.035 U/L, the PLP conjugated GSH_Mn_ZnS QDs was applied to quantify ALP activity in human serum and plasma. Further, the developed nanoprobe PLP conjugated GSH_Mn_ZnS QDs was also applied to detect Al3+. The complexation-induced fluorescence enhancement was observed at 492nm upon the interaction of Al3+ with the PLP conjugated GSH_Mn_ZnS QDs. Without any interference from other tested metal ions, this nanoprobe can be employed to detect Al3+ down to 2.30µM.

Anticalcification effects of DS-1211 in pseudoxanthoma elasticum mouse models and the role of tissue-nonspecific alkaline phosphatase in ABCC6-deficient ectopic calcification

Pseudoxanthoma elasticum (PXE) is a multisystem, genetic, ectopic mineralization disorder with no effective treatment. Inhibition of tissue-nonspecific alkaline phosphatase (TNAP) may prevent ectopic soft tissue calcification by increasing endogenous pyrophosphate (PPi). This study evaluated the anticalcification effects of DS-1211, an orally administered, potent, and highly selective small molecule TNAP inhibitor, in mouse models of PXE. Calcium content in vibrissae was measured in KK/HlJ and ABCC6-/- mice after DS-1211 administration for 13–14 weeks. Pharmacokinetic and pharmacodynamic effects of DS-1211 were evaluated, including plasma alkaline phosphatase (ALP) activity and biomarker changes in PPi and pyridoxal-phosphate (PLP). Anticalcification effects of DS-1211 through TNAP inhibition were further evaluated in ABCC6-/- mice with genetically reduced TNAP activity, ABCC6-/-/TNAP+/+ and ABCC6-/-/TNAP+/-. In KK/HlJ and ABCC6-/- mouse models, DS-1211 inhibited plasma ALP activity in a dose-dependent manner and prevented progression of ectopic calcification compared with vehicle-treated mice. Plasma PPi and PLP increased dose-dependently with DS-1211 in ABCC6-/- mice. Mice with ABCC6-/-/TNAP+/- phenotype had significantly less calcification and higher plasma PPi and PLP than ABCC6-/-/TNAP+/+ mice. TNAP plays an active role in pathomechanistic pathways of dysregulated calcification, demonstrated by reduced ectopic calcification in mice with lower TNAP activity. DS-1211 may be a potential therapeutic drug for PXE.